Investigation of Pomegranate (Punica granatum L.) Flowers’ Antioxidant Properties and Antibacterial Activities against Different Staphylococcus Species Associated with Bovine Mastitis

Abstract

Mastitis is one of the most considerable and costly diseases for dairy herds, and Staphylococcus spp. is known to be the main causative agent. Although antibiotics are widely used in the treatment of mastitis, this can cause both antibiotic residues in milk and the risk of antibiotic resistance occurrence in bacteria. Thus, in recent years, researchers have focused on alternative treatments for this disease and plants extracts are investigated for this purpose. Pomegranate is widely used as a dye, ornament, and medicinal plants in the industry, and the species has a particularly high economic value in Turkey. This study aims to investigate in vitro the antioxidant and antibacterial properties of the pomegranate flower’s extracts against different Staphylococcus species associated with bovine mastitis. To this aim, pomegranate flowers were collected from different regions in Turkey and extracts were prepared with three different solvents (methanol, ethanol, and water). The retention factor values of the ethanol extract were determined by thin-layer chromatography. The antibacterial activity tests were carried out via the disk diffusion method. In addition, the extracts were tested against the stable DPPH (2,2-diphenyl-1-picryl-hydrazyl-hydrate) free radicals for antioxidant activity. Four retention factors (0.79, 0.67, 0.58, and 0.33 points) were found for the ethanol extract. The methanol extract showed the highest inhibition zones against coagulase-negative Staphylococcus-37 (CNS-37) and S. aureus-18. The lowest MIC was 6500 μg/mL. The highest antioxidant activity was observed in methanol extracts. As a result, the extracts of pomegranate flowers showed a high antioxidant and antibacterial potential against the examined mastitis pathogens.

The mechanistic role of natural antimicrobials in preventing Staphylococcus aureus invasion of MAC-T cells using an in vitro mastitis model

“…Ajose et al 2023, have recently reviewed ethnoveterinary medicinal plants and ethnoveterinary medicinal products utilised in dairy and livestock farming to combat mastitis, with the most prominent plants as Becium obovatum , Malva parviflora , Brucea antidysenterica , Acorus calamus L., Triticum sp, Arachis hypogeam , Peganum harmala , Citrus limon , Withania somnifera and others [ 8 ]. Among plant extracts, Sorghum phenol extract [ 5 ], sweet orange peel ( Citrus sinensis ), pomegranate ( Punica granatum ) [ 22 , 23 ], indian screw tree ( Helicteres isora ) [ 24 ], lemon balm ( Melissa officinalis ), a specie of laurel wood ( Knema retusa ) [ 25 ] and olive [ 13 ] extracts demonstrated strong antimicrobial activity against different S. aureus strains including other BM-associated pathogens. After in vitro disc diffusion assays from recent studies, the authors have reported the inhibitory zones against S. aureus significant average zones ranging from 22 to 31 mm [ 24 , 26 ].…”

Evaluation of the Effects of Punica granatum Flower Tablets on Pain and Healing of Minor Recurrent Aphthous Stomatitis: A Randomized Clinical Trial

“…pleniflora decreased the time of healing more than others due to a higher amount of phenolic compounds, resulting in higher antioxidant activity. P. granatum flower has a variety of secondary metabolites such as polyphenols with strong antioxidative activity (Ökmen et al 2023 ). The polyphenols in P. granatum flowers contain alginic acid.…”

Polyphenol-Rich Extracts from Agroindustrial Waste and Byproducts: Results and Perspectives According to the Green Chemistry and Circular Economy

“…Antioxidant and anti-inflammatory effects are highlighted, resulting in a stabilization of the erythrocyte membrane, decreasing the quantity of hemoglobin derived from hemolysis. The flower extract inhibited nitrite generation and tumor necrosis (TNF-α) and showed activity against induced ischemia and brain damage in rats . A recent study on Punica granatum L. flower extract demonstrated the antioxidant and antibacterial activity against mastitis pathogens by reducing postinfection oxidative stress, highlighting the use of this natural extract for the treatment of this pathology.…”

“…In vivo Punica granatum leaves extract showed activity against ischemia-induced brain damage in rats that were treated with doses of 200–400 mg/kg for 7 days. This treatment reduced the brain changes that cause ischemia (edema, vascular congestion, and release of proinflammatory proteins and cytokines) . The study by Bekir et al.…”

“…Pomegranate leaves extracts, also rich in pigments, steroids, and terpenoids, have shown the ability to penetrate the microbial membrane and cause cell lysis. These effects have been demonstrated on strains of Pseudomonas aeruginosa , Escherichia coli , Salmonella spp., Staphylococcus aureus , Listeria monocytogenes , Enterococcus faecalis , Bacillus cereus , Candida albicans , and Aspergillus niger . Regarding pomegranate peels, a review by Singh et al collected in vivo and in vitro studies on the antibacterial and antifungal activity of pomegranate peel extracts rich in punicalagins, punicalins, gallic acid, ellagic acid, and gallic acid.…”